REPLIGEN KrosFlo TFDF User Manual Download

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KrosFlo® TFDF™ Lab System

User Guide

repligen.com IF.UG.023 R1

Pump priming process

Press

Manual

Press

Overview

Toggle Permeate pump P-02 to

FWD

Button will show green

Press

System Settings

Select

peristaltic tubing size

Example with #13 shown

Press

Overview

Press Permeate pump

P-02 flow rate

Input

flow rate value

in mL/min

Press

ENT

Press P-02

Start

Process graphic will flash green

Summary of Contents for KrosFlo TFDF

Page 1: ...1 IF UG 023 R1 KrosFlo TFDF Lab System User Guide ...

Page 2: ...dental or consequential damages in connection with the furnishing performance or use of this material No part of this document may be photocopied reproduced or translated to another language without the prior written consent of Repligen Corporation For further information please contact Repligen Corporation at www repligen com 2020 Repligen Corporation All rights reserved The trademarks mentioned ...

Page 3: ... process 27 Changing the default tubing calibration factors 35 Changing the system serial number 35 Updating the permeate flow meter installation status 36 Setting tubing sizes 36 Scaling 37 Changing the tubing calibration factors for a run not default 37 Resetting Tubing Calibration factors 37 Setting the maximum magnetic levitating recirculation feed pump P 01 speed 38 Setting pressure units 38 ...

Page 4: ... System User Guide KrosFlo TFDF Lab System User Guide 4 IF UG 023 R1 Experimental data 44 Run Setpoints screen 46 Overview screen 49 Instrumentation 56 Taring 57 Pump Speed 63 Pressure Flow 64 Weights 65 Data 66 PID 67 ...

Page 5: ...the bench 14 Figure 8 Assembling the stand 15 Figure 9 System connections 16 Figure 10 ProConnex TFDF Flow Path configuration 17 Figure 11 Connecting pressure sensors 21 Figure 12 Making tubing connections 22 Figure 13 Touchscreen display 23 Figure 14 Main Menu screen 24 Figure 15 Menu bar 24 Figure 16 Screen navigation 25 Figure 17 Set up before priming 26 Figure 18 Primed system 28 Figure 19 Exp...

Page 6: ...2 Initial screen 53 Figure 43 Initial Wizard feature screen 54 Figure 44 Calculation in progress 54 Figure 45 Manual Mode Overview screen 55 Figure 46 Reset prompt 56 Figure 47 Manual Mode Overview screen 57 Figure 48 Manual Mode Run Setpoints screen 58 Figure 49 Filter settings 58 Figure 50 Filter List screen left side 59 Figure 51 Navigation menu 59 Figure 52 Filter List screen right side 60 Fig...

Page 7: ...or cm Centimeter FAS Field Application Scientist Hz Hertz in Inches kg Kilogram lbs Pounds LMH Liters Meters2 Hour LPM Liters per minute PCV Percent Cell Volume PE Pressure Sensor PID Proportional Integral and Derivative PSI Pounds per square inch PV Process variable rpm Reps per minute SP Desired setpoint TFDF Tangential Flow Depth Filtration TMP Transmembrane Pressure UL Underwriters Laboratorie...

Page 8: ...ument for your KrosFlo TFDF Lab System and is updated on a regular basis For the latest version of the document please visit www repligen com resources It is highly recommended that the installation process be executed by a trained Repligen engineer For further support with troubleshooting or process optimization please contact your local Repligen Field Application Scientist Welcome Thank you for ...

Page 9: ...or nature of hazard and corrective actions Potentially hazardous situation which if not avoided may result in property equipment damage Caution Risk of crushing Keep fingers away from rotor while pump is in operation Stop pump before loading or unloading tubing Caution Hot surface Do not touch Caution Risk of electric shock consult Operating Instructions for nature of hazard and corrective actions...

Page 10: ...al No foreign matter should be allowed under the gasket on the back of the front plate or under the heads of the screws Failure to observe this precaution may result in leakage during washdown of the drive Caution To avoid electrical shock the power cord protective grounding conductor must be connected to the ground Not for operation in wet locations as defined by EN61010 1 Warning High impeller m...

Page 11: ...tation The Pump Station transfers all liquids during the TFDF process A low shear and digitally controlled magnetic levitating centrifugal style pump delivers feed stock from the bioreactor vertically through the lumen of the TFDF filter and then back into the bioreactor The Permeate top peristaltic pump transfers permeate from the filter enclosure to the permeate reservoir The Diafiltration Aux p...

Page 12: ... Stand mount 6 Flow meter 7 Control panel 8 Tubing guide rod 9 Extension rod 10 Sleeve 11 Filter clamp TFDF Filter The TFDF Filter is a tubular depth filter available in process development to pilot and production scales The filter functions within an enclosure with ports that deliver feed stock and remove retentate and permeate All filter sizes larger than the trial size are available as ProConne...

Page 13: ...k genderless aseptic connector 2 Retentate tubing 3 Retentate pressure sensor PE02 4 Vent port 5 Pinch clamp 6 TFDF filter enclosure 7 Permeate pressure sensor PE03 8 Feed pressure sensor PE01 9 CPC AseptiQuik genderless aseptic connector 10 Magnetic pump head 11 Ferromagnetic fixation disc 1 9 2 3 4 5 6 7 8 10 11 ...

Page 14: ...e are recommended to lift the Controller out of the box and place it on the bench top FRONT VIEW SIDE VIEW 5 ft 150 cm Set up Space requirements Figure 6 Bench space needed System arrangement Figure 7 Arranging the system on the bench 1 Controller 2 Pump Station 3 Scale for bioreactor 4 Scale for permeate 1 2 3 4 ...

Page 15: ...epligen com IF UG 023 R1 Stand assembly Figure 8 Assembling the stand 5 Tubing guide rod 6 Extension rod required only for 108 cm flow path 7 Filter clamp 8 Sleeve 20 cm flow path configuration 108 cm flow path configuration 5 6 8 7 5 6 8 7 ...

Page 16: ...nnections 1 Connect the 718 Minifast Turbidity sensor cable from the Pump Station to your turbidity sensor 2 Connect the longer M12 Eurofast depth station flow meter cable from the Pump Station to your depth station KEY Required Optional Non cable Note Use tear drop to align pins with socket Communication cable pins are delicate ...

Page 17: ...ure 10 ProConnex TFDF Flow Path configuration 1 CPC AseptiQuik genderless aseptic connector 2 Retentate tubing 3 Retentate pressure sensor PE02 4 Vent port 5 Pinch clamp 6 TFDF Filter enclosure 7 Permeate pressure sensor PE03 8 Feed pressure sensor PE01 9 CPC AseptiQuik genderless aseptic connector 10 Magnetic pump head 11 Ferromagnetic fixation disc 1 9 2 3 4 5 6 7 8 10 11 Example ProConnex TFDF ...

Page 18: ...stall flow path Unbox the flow path and remove the ferromagnetic disc 20 cm flow path installation 1 2 3 Align pump head Pull locking pin and insert pump head Rotate filter up locking pin will click Note Remove the ferromagnetic disc before proceeding to the next step ...

Page 19: ...od with tubing guide rod to sleeve and rotate clamp so that it is out of the way during installation Connect retentate pressure sensor with RJ12 extender to PE02 This is much easier to connect before the flow path is installed due to its height 1 2 3 Note RJ12 cables are not supplied with the ProConnex assemblies and must be purchased separately Note Extender available from Repligen support ...

Page 20: ... IF UG 023 R1 Align pump head Pull locking pin and insert pump head Rotate filter up locking pin will click 7 Secure flow path with clamp and raise extension rod so that guide rod is at retentate tubing height Make final adjustments as necessary 4 5 6 ...

Page 21: ...ors Connect pressure sensors PE01 to Feed PE02 to Retentate for 108 cm flow path already connected in Installation step 3 PE03 to Permeate PE04 to Secondary filter OPTIONAL PE05 to Guard Sterile filter OPTIONAL PE06 NOT USED Connect in line turbidity sensor to permeate line OPTIONAL Note In line turbidity sensor cables are not supplied with the system and must be purchased separately ...

Page 22: ...ubing connections Make tubing connections 1 Route retentate tubing over tubing guide stand adjustments may be required 2 Route permeate tubing through top peristaltic pump 3 Route diafiltration buffer tubing through bottom peristaltic pump 4 Feed line not installed in flow meter enables easier priming ...

Page 23: ... started Initial startup Turn the KrosFlo TFDF Lab System on using the power switch located on the left rear panel of the main enclosure Once the system has booted up the information screen shown below will display Touch the screen to continue Figure 13 Touchscreen display ...

Page 24: ...reen provide access to different operational and set up screens To navigate to a different screen simply touch the button Screen navigation The menu bar displays at the top of all system screens Figure 15 Menu bar Screen navigation options display on the bottom of all system screens The options shown vary from screen to screen Set alarms and view history Name of screen currently displayed Locks th...

Page 25: ...en navigation Screen saver The system is programmed with a screen saver that comes on after 30 minutes of inactivity This does not affect operation in any way When the screen saver is active the display will be black Just touch the display screen to view the active system screen ...

Page 26: ...nd into the magnetic levitation pump Priming is typically not required for bottom fed bioreactors Pump priming set up Figure 17 Set up before priming Verify the following set up steps have been executed prior to pump priming Feed pressure sensor connected to PE01 Retentate pressure sensor connected to PE02 Permeate pressure sensor connected to PE03 Vent line clamped closed Retentate line clamped c...

Page 27: ...l Press Overview 2 Toggle Permeate pump P 02 to FWD Button will show green Press System Settings 3 Select peristaltic tubing size Example with 13 shown Press Overview 4 Press Permeate pump P 02 flow rate 5 Input flow rate value in mL min Press ENT 6 Press P 02 Start Process graphic will flash green ...

Page 28: ...to the feed line and pump chamber Stop the peristaltic permeate pump P 02 when fluid can be seen in the feed pressure sensor above the pump chamber 8 Open clamp on retentate line 9 Route feed line through flow meter Priming is complete Figure 18 Primed system IMPORTANT It is critical that no fluid enters into the TFDF Filter element during priming ...

Page 29: ...e direction At the end of Step 1 approximately 50 of the original cell culture volume has been transferred to the permeate reservoir and the cell culture feed has been effectively concentrated Step 2 Wash Diafiltration Wash buffer purple is pumped into the bioreactor while circulation of the cell culture feed stock continues Permeate continues to accumulate in the permeate reservoir At the end of ...

Page 30: ...reservoir At the end of Step 3 clarification of the initial bioreactor feed stock had completed and the permeate reservoir volume had reached approximately 110 120 of the initial cell culture feed volume Completion Measure turbidity and product concentration in the permeate reservoir to calculate turbidity reduction and yield Disconnect the permeate reservoir and store for the next operation Dispo...

Page 31: ...ine TFDF turbidity reduction Crossflow rate Fixed value at 2L min fiber Target yield Typically 90 95 Targeting higher yields may increase permeate turbidity Targeting higher yields may increase diafiltration buffer requirement Maximum final pool volume Typically 110 120 of starting cell culture feed volume Increasing may increase yield with challenging samples Initial concentration factor Fold con...

Page 32: ... an initial dilution of the feed material in buffer or media Most samples meet recovery and turbidity reduction expectations with Wizard feature derived parameters and without optimization If turbidity breakthrough is observed it is recommended to increase volumetric expansion by 5 10 and reduce the initial concentration factor If filter fouling is observed it is recommended to verify the PCV meas...

Page 33: ...rview Shows all the available auxiliary inputs aux pumps pressure sensors scales flow meters turbidity meter and their live values available options on the screen will change depending on what automation mode is selected Mode Allows the user to select an automation mode and select a filter part number Run Allows the user to input run setpoints parameters and use the Wizard feature Plots Shows live...

Page 34: ... 21 Main Menu screen Admin screen The Admin screen allows users to adjust default tubing calibration factors change the system serial number and update the permeate flow meter installation status To access this screen select the Admin Screen button in the Main Menu screen ...

Page 35: ...nging default values is done at the Admin level only To change a tubing calibration factor setting select one of the blue fields and enter a new default value Setting the calibration factor here sets the default for operation Users can restore tubing calibration factors to default values using the Reset Tubing Calibration button in the System Settings screen For more information see the System Set...

Page 36: ...sizes and calibration factors product vessel capacity and pump speeds To access this screen select the System Settings button in the Main Menu screen Figure 23 System Settings screen Setting tubing sizes 1 Confirm the size of the tubing that will be used by noting the size printed directly on the tubing itself 2 Select a circle button under the P 02 or P 03 Tube Size column to view size options Th...

Page 37: ...n not default 1 Select a grey box next to a tubing size under the Pump P 02 or Pump P 03 2 Choose a flow rate rpm and measure the output on a scale 3 Add the rpm ml number in the respective box for calibration The following table shows the available flow rates for the KrosFlo TFDF Lab system Table 6 KrosFlo TFDF Lab System permeate tubing flow rates Tubing size Low range ml min High range ml min 1...

Page 38: ...king into account the gain factor K Using proportional control alone will result in an error between the setpoint and the actual process value because it requires an error to generate the proportional response If there is no error there is no corrective response 2 Term I accounts for past values of the SP PV error and integrates them over time to produce the I term For example if there is a residu...

Page 39: ...m response To access this screen select the PID Set up button in the Main Menu screen Values for Proportional Integral and or Derivative loop settings can be added for Recirc Feed pump control Reactor volume control and Permeate flow control The default values are optimized for stable control and are recommended To add or change values select a blue box and enter a value Figure 25 PID loop setting...

Page 40: ... on the tubing exiting the filter is increased Check for bent tubing or obstructions in the tubing lines back to the recirculation vessel 3 High Permeate Pressure PE 03 Monitors if the permeate pressure on the filtrate side is high due to an obstruction or bent tubing 4 Low Permeate Pressure PE 03 Indicates a fouled filter Terminate the run if near completion or reduce the flux to finish the run 5...

Page 41: ...e full system For example the recirculation feed pump will stay on the permeate pump will stop and the diafiltration pump will stop to maximize the opportunity to recover the run High alarms will trigger when the process value rises to the stored setpoint or above Low alarms will trigger when the process value falls to the stored setpoint or below The system is equipped with a short delay to avoid...

Page 42: ...select Alarms in the menu bar To erase the list of prior alarms select Clear History button Figure 28 Alarm History screen Lock screen The Lock option on the KrosFlo TFDF System allows users to lock the screen for cleaning without inadvertently affecting system operation 1 Select Lock in the menu bar The following message will display Figure 29 Lock screen message 2 Select Yes The Lock Screen will...

Page 43: ...m to a USB drive To access this screen select Data Log from the Main Menu screen Figure 31 Data logging screen To record process data insert a USB drive into the USB port of the Controller Data logging activates automatically with detection of a USB drive When the data is logging the USB Ready button will be green and display On When the data is writing the USB Writing button will be green display...

Page 44: ...Permeate o PE04 Secondary Filter o PE05 Sterile Guard Filter TMP Calculated Feed pressure Retentate pressure 2 Permeate pressure Scale readings kg o Feed scale o Retentate scale Module surface area calculated from selected filter PN Fiber Count Pi Effective Length Fiber size Flow meters o Feed flow measurement o Permeate flow measurement optional o Permeate totalizer optional calculated using the ...

Page 45: ...oncentration Diafiltration Mode Automated filtration mode where retentate is concentrated to a certain concentration factor then run in diafiltration mode where retentate volume is kept constant through addition of buffer media 3 Concentration Diafiltration Concentration Mode Automated filtration mode where retentate is concentrated to a certain concentration factor then run in diafiltration mode ...

Page 46: ...ation of select setpoints or options for the operational mode To access this screen select the Run button at the bottom of the System Mode Screen Concentration Mode Run Setpoints screen used to configure a simple draw down process Users can select the Concentration Factor CF or Permeate Weight as the setpoint Figure 35 Concentration Mode Run Setpoints screen Concentration Diafiltration Mode Run Se...

Page 47: ...ts Screen is used to configure an initial drawdown buffer addition and a final drawdown users have the same options for setpoints as the previous modes selecting the Start Concentration Diafiltration Concentration button in this mode initiates the Wizard feature that automatically runs calculations to determine various setpoints for more information refer to the Wizard feature section ...

Page 48: ...trols the rpm of the permeate pump to match target permeate rate Initial Concentration SP CF1 A dimensionless concentration factor used to quantify concentration of the feedstock it is the amount that the feedstock has been reduced in volume relative to the initial volume for example if 1 L of feedstock is concentrated to 0 25L with 0 75 L padding through the filter as permeate a 4 fold concentrat...

Page 49: ...tial starting volume not from the start of the second concentration for instance if 1 L of feedstock is processed until 0 75 L has passed through to the filtrate and 0 25 L is left in the retentate a 4 fold concentration has been performed so the Concentration Factor is 4X and the input would be 4 Permeate Weight This value refers to the total cumulative permeate weight at end of the final concent...

Page 50: ...ode operations User input of Concentration Factor or Permeate weight for Concentration step User input of Diafiltration Volume or Permeate weight for Diafiltration step Change direction of Auxiliary pumps P 02 and P 03 Tare scales WE 01 and WE 02 Tare pressure sensors PE 01 PE 02 PE 03 PE 04 PE 05 and PE 06 Zero Flow meter FL 01 and FL 02 ...

Page 51: ...f Concentration Factor or Permeate weight for Concentration 1 step User input of Diafiltration Volume or Permeate weight for Diafiltration step User input of Concentration Factor or Permeate weight for Concentration 2 step Change direction of Auxiliary pumps P 02 and P 03 Tare scales WE 01 and WE 02 Tare pressure sensors PE 01 PE 02 PE 03 PE 04 PE 05 and PE 06 Tare Flow meter FL 01 and FL 02 ...

Page 52: ... Cell Volume Initial Concentration Factor CF 1 Expected Final Yield Final Permeate Volume The Wizard feature is accessed in the Run Setpoints screen when the Concentration Diafiltration Concentration mode is selected Its calculations determine Diafiltration buffer volume needed Permeate volume at diafiltration pump start Permeate volume at diafiltration pump stop Permeate volume at run end Time fo...

Page 53: ...ll display Figure 42 Initial screen 2 The Wizard feature initially displays default values To enter a value and perform a calculation select a settings button and enter the appropriate value Repeat this step for any other settings that need to be calculated Note Users can enter values for one multiple or all settings in the Wizard feature ...

Page 54: ... Calculation successful message will display once the calculation is complete If a Calculation Failure message displays valid setpoint criteria could not be determined based on the input Adjust the entered values and restart the calculation If a Calculation Timeout message displays select RESET and restart the calculation To exit the Wizard feature select Close Select a button to enter a value Sta...

Page 55: ... flow meter FL 01 for the feed pump P 01 control or scale WE 01 for the Aux Diafiltration pump P 03 control Figure 45 Manual Mode Overview screen Any combination of running auxiliary components is possible in Manual Mode there are no set points to automate Start stop main pump P 01 Start stop auxiliary pumps P 02 and P 03 Change direction of auxiliary pumps P 02 and P 03 Tare scales WE 01 and WE 0...

Page 56: ...ear rate at the wall of the fiber based on the number of fibers fiber ID and recirculation flow rate Instrumentation P 01 Magnetic Levitating Recirculation Feed Pump P 02 Permeate Pump top peristaltic pump on Pump Station P 03 Diafiltration Aux Pump bottom peristaltic pump on Pump Station For more detailed instructions about the functionality of the overview screens please refer to the System Mode...

Page 57: ...verview screen and described in their respective mode sections Flow meter Zero Zeros the flow meter located between the bioreactor and the filter zero the flow meter only after flow path is primed only the zero button disappears during automation sequences to prevent accidental zeroing Reset Totalizer While the permeate is flowing the totalizer sums the total volume For accuracy reset the totalize...

Page 58: ... settings Select Module displays the filter modules currently available and lets users select the filter module best suited for their application Part Number displays the part number for the currently selected filter module Surface Area displays the membrane surface area as calculated by inner circumference multiplied by the length of the fiber To view available filter modules and select one for t...

Page 59: ...y Figure 51 Navigation menu Select the and magnifying glass icons to zoom in and out Select the arrows to scroll left right or up down in the list Select the white grey box to change the table view between alternating grey and white rows alternating grey and white columns or all white The initial table displays the Fiber Size and Fiber Count columns To view EFF Length select the Navigation icon an...

Page 60: ... To select a specific filter module 1 Select a row which is associated with its respective part number for the desired filter family 2 Select the Navigation icon 3 Select the Load button 4 Select Close The part number for the filter module selected will now display in the filter options in the System Mode Screen and will be used during system operation ...

Page 61: ...storical data using the forward and reverse arrows at the top left side of the screen Each plot has a set of pens shown in the pen toolbar at the bottom of the screen each pen represents data for a specific system parameter and has a unique trace color in the plot to remove a parameter from the plot display toggle the eyeball button next to the parameter see Table 13 for a list of all parameters p...

Page 62: ...Feed flow rate LPM FL 02 Permeate flow ml min WE 01 Product vessel weight kg WE 02 Permeate vessel weight kg PE 01 Feed pressure Psig or mbar PE 02 Retentate pressure Psig or mbar PE 03 Permeate pressure Psig or mbar P 01 P 02 P 03 Feed pump set point Pump speed P 02 Pump speed P 03 RPM or LPM RPM or ml min RPM or ml min ...

Page 63: ...ide 63 repligen com IF UG 023 R1 Pump Speed Pen traces in the Pump Speed plot display trend data for the following P 01 PID set point P 01 PID PV P 02 PID set point P 02 PID PV P 03 PID set point P 03 PD PV Figure 54 Pump Speed plot ...

Page 64: ...Flow Pen traces in the Pressure Flow plot display trend data for the following PE 01 Feed pressure PE 02 Retentate pressure PE 03 Permeate pressure Recirc Feed flow Permeate flow PE 04 Pre sterile filter PE 05 Post sterile filter FL 02 Scale flow Figure 55 Pressure Flow plot ...

Page 65: ...sFlo TFDF Lab System User Guide 65 repligen com IF UG 023 R1 Weights Pen traces in the Weights plot display trend data for the following Reactor weight Permeate weight Permeate total Figure 56 Weights plot ...

Page 66: ...ay trend data for all tracked parameters PE 01 Feed pressure PE 02 Retentate pressure PE 03 Permeate pressure Calculated TMP PE 04 Pre sterile filter pressure PE 05 Post sterile filter pressure Feed Weight Permeate Weight Surface area Recirc Feed flow Permeate total FL 02 Scale flow Figure 57 Data plot ...

Page 67: ...tem User Guide 67 repligen com IF UG 023 R1 PID Pen traces in the PID plot display trend data for the following P 01 PID set point P 01 PID PV P 02 PID set point P 02 PID PV P 03 PID set point P 03 PD PV Figure 58 PID plot ...

Page 68: ...seated properly in the magnetic levitating pump 3 Inspect the pump head for clogs 4 Inspect the filter for clogs Pump Station not responding There are several different communications protocols for the Pump Station To address most issues shut the system down then disconnect and reconnect the 26 pin communications cable from the Pump Station to the Main Enclosure see connect system cables sections ...

Page 69: ...uct warranty The KrosFlo TFDF Lab System is manufactured in Marlborough MA USA General information Safety guidelines Figure 59 Magnetic forces caution and warning Table 8 Warning Product use limitation Symbol Description Caution Risk of danger Consult Operating Instructions for nature of hazard and corrective actions This product is not designed for nor intended for use in patient connected applic...

Page 70: ...tate flow meter Ultrasonic clamp on flow meter 0 8000 ml min 2 accuracy 16 ml min Calibrated for 15 PharmaPure tubing Recommended process volume 1 50 L Number of pressure sensors supported 5 Pressure sensor range 14 30 psi 1 2 bar TFDF Filter surface area supported 2 150 cm2 Number of scales supported 2 Table 10 System input Description Specifications Power requirements 120 VAC 10 A 230 VAC 5 A 50...

Page 71: ...e polycarbonate polysulfone and C Flex PharmaPure materials System components Table 13 System components list Item Included Components Controller Controller with housing connected cables Pump Station Pump Station attachments o Peristaltic pumps x2 o Magnetic levitation pump o Stand mount with locking knob o Flow meter A C Power cables US UK EU and China versions included Pump Station power cable 5...

Page 72: ...afiltration 7 12 13 32 45 49 50 52 53 54 55 58 59 73 Flow path 13 16 19 20 22 52 60 Installation 8 19 20 35 37 LMH 7 32 Mode 32 45 47 48 49 50 52 55 60 61 Precautions 9 Pressure sensor 13 14 18 22 27 60 ProConnex 13 14 18 Pump control 40 Pump Station 12 13 15 17 59 71 73 74 Requirements 15 73 Safety 9 72 Scale 38 45 50 58 Set up 25 27 34 Shear 12 42 59 Specifications 12 18 73 TMP 7 45 Warning 9 42...

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